tbuckley/cabot-search huggingface dataset:
PB2179: ALGORITM OF MOLECULAR GENETIC EXAMINATION OF PATIENTS WITH PH-NEGATIVE CRONIC MYELOPROLIFERATIVE NEOPLASM
Authors: Myroslav Voroniak, Zvenyslava Maslyak, Maryana Kokoruz, Sofiia Hudzij, Nataliya Shurko
Journal: HemaSphere (2023)

Topic: 15. Myeloproliferative neoplasms - Biology & Translational Research Background: Molecular biology studies were identified mutations in the genes that trigger the pathological process in Polycythemia Vera (PV), Essential Thrombocythemia (ET) and Primary Myelofibrosis (PMF): JAK2, MPL and CALR. These “driver” mutations underlining their role in the emergence of the myeloproliferative process, its clinical features, the development of complications and therapeutic tactics in some Ph-negative Chronic Myeloproliferative Neoplasms (MPN Ph-). The V617F mutation is most often found in exon 14 of the JAK2 gene (80-90% of patients with PV and 50-60% of patients with ET or PMF). Another 4% of patients with PV identify mutations in exon 12 of the JAK2 gene. 67-88% of JAK2-negative patients with ET and PMF show mutations in the CALR gene. The mutations of the MPL W515 gene in MPN are the rarest and the most cases are associated with the W515K and W515L mutations. These mutations are found in 3-8% of PMF and ET cases. However, 5-10% of patients with ET and PMF are “triple negative” (without mutations in the JAK2, MPL and CALR genes). Aims: to create an algorithm for the molecular genetic examination of NMPC Ph- patients. Methods: Peripheral blood samples were used in patients with suspected or/and diagnosed MPN Ph-. Blood samples were taken on an empty stomach in tubes with 6 % ethylenediaminetetraacetic acid (EDTA). The study was performed with DNA isolated from peripheral blood leukocytes. Amplification was performed on a CFX96TM Real-Time System (Bio-Rad (USA)). Results: Mutation studies were conducted in 181 patients with NMPC Ph-. The JAK2 V617F mutation was determined in 36% patients. Among the JAK2-negative patients, 8% had type 1 mutations in the CALR gene (52 bp deletions), 4% had type 2 mutations in the CALR gene (5 bp insertions), and 2% had mutations in W515K and W515L. A three-stage algorithm of molecular genetic examination of MPN patients has been proposed. In the first stage of diagnosis, the JAK2 V617F mutation was determined and the patients were examined using other diagnostic methods (histological, cytological and cytogenetic). In JAK2-negative patients, the next step was to identify mutations in the CALR gene and, in case of a negative result, the third step was to detect W515K/L mutations in the MPL gene. Summary/Conclusion: Implementation of the proposed study scheme in clinical practice will help improve the diagnosis of patients with suspected MPN Ph-, as well as provide adequate treatment tactics in patients based on their molecular genetic characteristics. Keywords: Myeloproliferative disorder
PB2364: DEMOGRAPHICS, PATTERN OF CARE, AND OUTCOMES OF PRIMARY NODAL PERIPHERAL T-CELL LYMPHOMAS. DATA FROM THE MOLDOVAN HEMATOLOGICAL CENTER.
Authors: Victor Tomacinschii, Maria Robu, Sanda Buruiana
Journal: HemaSphere (2023)

Topic: 19. Aggressive Non-Hodgkin lymphoma - Clinical Background: Peripheral nodal T-cell lymphoma (PTCL-N) is a subtype of peripheral T-cell lymphoma (PTCL) characterized by the involvement of lymph nodes by malignant T cells. PTCL-N are a heterogeneous group of diseases that can have varying clinical, pathological, and genetic features. PTCL-N accounts for approximately 10-15% of all non-Hodgkin’s lymphomas (NHL) in Western countries. Aims: To investigate the clinicopathological characteristics of patients with PTCL-N and review their outcomes features. Methods: We performed an observational study on a group of patients with PTCL-N diagnosed during 2017-2022 at the Oncological Institute of Moldova. During this period, 22 patients with PTCL-N were diagnosed and monitored in the Hematology clinic of the Moldovan Oncology Institute. Results: The median age was 52,19% years (range 25-84) and 54,54% were female. Most of the patients were diagnosed in generalized stages: stage III - 4 (18,18%) cases, stage IV in 13 (59,09%) patients. Localized stages were found in 22,72% of cases, of which: stage I - 1 (4,54%) patients, stage II - 4 (18,18%) cases. Among the patients with IV stage at the diagnosis, 61.53% had evidence of extranodal disease. Considering the IPI score, patients with high risk have been identified 45,45% of cases; The primary affected area developed in 45.45% in the cervical lymph nodes, in 36.36% in the axillary lymph nodes, and in 18.18% the onset was associated with the involvement of the inguinal nodes. The predominant morphological form was PTCL NOS determined in 12 (54.54%) cases, followed by Anaplastic T cell lymphoma ALK-positive, 4 (18.18%), and Anaplastic T cell lymphoma ALK-negative in 2 (9.09%). Angioimmunoblastic T cell lymphoma was identified in 3 (13.63%). PTCL unspecified morphological type has been found in 1 case (4.54%). 90,9% of the patients received protocols including anthracyclines such as CHOP in 65% (n=13) or CHOEP in 35% (n=7) as first-line treatment. Following first-line treatment, complete remissions were determined in 54.5% of cases, partial remissions in 27.2%, progressive disease was determined in 18.1%. Overall 3-year survival rate was 40.9%. Factors associated with the poor outcomes that have been found are age>65 years; high IPI score; increased LDH; Summary/Conclusion: According to our observations, the most common type PTCL-N that was diagnosed is PTCL-N NOS (54,54%). Majority of the patients have at the presentation a generalized stage (77,27%), extranodal involvement (61,53%), and high IPI score (45,45%). PTCL-N is a concerning diagnosis with a poor prognosis (OS at 3 years = 40,9%).Figure 1. Distribution of PTCL-N subtypes within the study group Keywords: Prognosis, T cell lymphoma, Peripheral T-cell lymphoma, Chemotherapy
PB1703: MODELING T-CELL DEVELOPMENT AND T-ALL INITIATION
Authors: Advaita M Dick, Erika Tsingos, Narges Aghaallaei, Julia Skokowa, Baubak Bajoghli
Journal: HemaSphere (2023)

Topic: 1. Acute lymphoblastic leukemia - Biology & Translational Research Background: T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive blood malignancy derived from abnormal developing T-cells (thymocytes) within the thymus. In humans and mice, T-ALL is diagnosed when malignant T cells are detectable in the peripheral blood, i.e., at a stage when disease development is complete. Thus far, the ontogeny of T-ALL is unknown. Therefore, we are modeling T-cell development and disease onset. Aims: We aim to get a deeper understanding of T-ALL initiation and progression. Methods: We used an interdisciplinary approach to address this issue: First, we took advantage of our recently developed “virtual thymus”, a computational model of thymus organ (Aghaallaei et al., Science Advances, 2021), and simulated conditions in which either the level of cytokine interleukin (IL-7) by thymic epithelial cells (TECs) or its receptor on a thymocyte was upregulated. Additionally, we introduced a dominant active interleukin 7 receptor (IL-7R) mutation. The outcome of simulations was then tested with our animal model system by genetically engineered thymocytes and TECs. Results: In an unbiased strategy, we simulated >300 different conditions and selected scenarios that resulted in clonal expansion. Consistent with in silico analysis, our in vivo model showed that manipulation of thymocytes, either by IL-7, IL-7R or Notch1 upregulation, resulted in thymoma formation within less than 3 days only in 10%, 25% or 30% of cases, respectively. However, this frequency was accelerated to 80% when both the dominant active IL-7R level and Notch were upregulated. Summary/Conclusion: We offer new insights in the mechanism that may trigger T-ALL initiation in the thymus. In addition, we show the “virtual thymus” as a new functional in silico model to predict the outcome of various conditions for the T-cell and T-ALL development or treatment. Keywords: T-ALL, In vivo imaging, Animal model, Thymus
P1626: A SINGLE-CENTER EXPERIENCE IN INCREASING FACTOR VIII LEVELS TO ACHIEVE ZERO BLEEDING IN PATIENTS WITH SEVERE HEMOPHILIA A.
Authors: Rubén Clavero López, Francisco José López Jaime, Eva María Calavia Aranda, Ana María Mena Santano, Adrián Montaño, Ihosvany Fernández Bello
Journal: HemaSphere (2023)
P1251: LNCRNA NONHSAT064292.2 PROMOTES INTESTINAL AGVHD BY REGULATING AUTOPHAGY THROUGH MODULATING MIR-1225-3P AND TARGETING RPTOR
Authors: Jiaqi Fu, Xing Cui
Journal: HemaSphere (2023)

Topic: 21. Stem cell transplantation - Experimental Background: Intestinal acute graft-versus-host disease (aGVHD) is one of the major complications of allo-HSCT, manifesting with clinical signs of diarrhea, abdominal pain sensation, and hematuria1. Previous studies have confirmed that abnormal autophagy levels involve in the progression of intestinal aGVHD2,3. Meanwhile, the role of lncRNAs in peripheral blood mononuclear cells (PBMCs) in promoting aGVHD occurrence and development is also being revealed4,5. Therefore, we try to investigate the prognostic potential of lncRNA in aGVHD and whether it can influence aGVHD pathology by regulating autophagy. Aims: To explore the specific role of aberrant lnc-NONHSAT064292.2 in intestinal aGVHD pathogenesis. Methods: PBMCs were extracted from fifteen aGVHD patients and four healthy controls to generate the aberrantly expressed lncRNA profiles by high-throughput sequencing. PCR validation and correlation analysis were conducted between lncRNA and clinical characteristics of aGVHD. The ceRNA interaction network was verified by the dual-luciferase reporter assay. Caco-2 cells induced by TNF-α were established as the model for intestinal barrier dysfunction. The mice models with aGVHD were established by transplanting a mononuclear cell suspension from Balb/c donor mice treated with 60Co X-rays into CB6F1 recipient mice. Intestinal mucosal pathology was evaluated by HE staining. Immunofluorescence (IF), immunohistochemistry (IHC) and western blotting (WB) were used to detect expression of autophagy-related indicators (LC3II/I, Beclin 1 and P62). Transmission Electron Microscopy (TEM) was used to observe the number of autophagic vesicles. Results: Lnc-NONHSAT064292.2 with high expression in aGVHD was selected and confirmed by PCR (Figure A). Correlation analysis confirmed that the expression of lncRNA was strongly correlated with clinical characteristics of aGVHD (Figure B). Then, dual-luciferase reporter assay revealed that Lnc-NONHSAT064292.2 might induce aGVHD via the downstream miR-1225-3p and RPTOR (Figure C). The results of WB, IF and TEM confirmed that lncRNA could increase the RPTOR level to suppress autophagy through the ceRNA mechanism and promote intestinal aGVHD progression (Figure D-F). The intestinal mucosal histology of different groups indicated that intestinal aGVHD was alleviated when the autophagy level was induced (Figure G). The results of WB, TEM, and IHC indicated the RPTOR-related low-level autophagy in intestinal aGVHD (Figure H).Summary/Conclusion: Lnc-NONHSAT064292.2 could influence RPTOR-induced autophagic dysregulation by sponging miR-1225-3p in intestinal aGVHD. *Correspondence to: Xing Cui, M.D and Ph.D. Taishan Scholar, Professor. Keywords: Acute graft-versus-host disease
P1363: NOVEL DUAL-TARGETING AND SURFACE-ENGINEERED BIO-ORTHOGONALLY LENTIVIRAL VECTORS FOR IN VIVO CAR-T CELLS GENERATION
Authors: Zhaozhao Chen, Yu Hu, Heng Mei
Journal: HemaSphere (2023)

Topic: 24. Gene therapy, cellular immunotherapy and vaccination - Biology & Translational Research Background: Chimeric antigen receptor T-cell (CAR-T) therapy has emerged as markedly efficacious treatment modalities in recent years for refractory and relapsed hematopoietic malignancies. However, the exorbitant cost and intricate manufacture procedure of ex vivo generation has hampered its broader application. In vivo induced CAR-T therapy offers the potential to obviate cumbersome manufacturing logistics, providing new insights into CAR-T treatment. We developed a novel, dual-targeting and surface-engineered lentiviral vectors for reprogramming circulating T lymphocytes to CAR-T cells and eliminating tumor cells in vivo. Aims: This study aims at inducing CAR-T cells generation in vivo and wiping out tumor cells by novel lentiviral vectors. Methods: To engineer dual-targeting lentiviral vectors (D-LVs), the packing cell lines 293T was labelled with azide groups by glycometabolic bioorthogonal chemistry (designated N3-293T) firstly, and a CD3 promoter (CD3p) was fused to CAR transfer plasmid. Secondly, azide groups modified LVs were further surface engineered with anti-CD3 antibody (OKT3) via click chemistry. The conjunction of OKT3 and was analyzed using dot immunobinding assay, confocal microscopy and the in vitro transduction efficiency was evaluated using flow cytometry. To demonstrate the ability of targeted transduction and specific cytotoxicity in vivo, D-LVs were intravenously infused into humanized NOD-scid-IL2Rγnull (huNSG) mice engrafted with Nalm6-luc cells. Subsequently, the tumor burden was monitored using a noninvasive bioluminescence imaging system and the in-vivo CD19-CAR-T cells existence were detected by flow cytometry. Results: By displaying OKT3 on the single lentiviral surface and fused CD3p into the key construct, we could achieve targeted delivery of CD19-CAR genes to T cells both in vitro and in vivo. Summary/Conclusion: These results demonstrate the great potential applications of this engineered lentiviral system as a new strategy for inducing CAR-T immunotherapy in vivo and a promising approach for leading personalized treatment.Keywords: Immune therapy, CD19, CAR-T, T cell
P1562: THE FIRST TWO YEARS OF THE EUROPEAN VACCELERATE VOLUNTEER REGISTRY
Authors: Jon Salmanton-García, Fiona A. Stewart, Sarah Heringer, Janina Leckler, Oliver A. Cornely, Zoi Dorothea Pana
Journal: HemaSphere (2023)

Topic: 30. Infections in hematology (incl. supportive care/therapy) Background: The VACCELERATE Volunteer Registry is an active single-entry point for European residents interested in clinical trial participation. It has received funding from the European Union’s Horizon 2020 research and innovation programme (grant agreement 101037867), and the German Federal Ministry of Education and Research (grant BMBF01KX2040). As of today, the VACCELERATE Volunteer Registry is active in 17 countries, and associated to two national registries (France, The Netherlands), with 106,869 registered volunteers overall. Participants provide their contact information, including first and last name and e-mail address, age, gender, pre-existing comorbidities, vaccination status for SARS-CoV-2, COVID-19 history, and maximum distance willing to travel to a clinical trial site, if needed. The registry is open to both adults and children, complying with legal consent requirements. The VACCELERATE Volunteer Registry is active since October 2020. Aims: The VACCELERATE Volunteer Registry aims to connect all stakeholders in the clinical trial performance (volunteers, clinical trial sites, promoters…) to facilitate the clinical trial performance in Europe. Methods: Once volunteers sign in on www.vaccelerate.eu/volunteer-registry, data are stored until a request is received from entities managing or performing clinical trials or epidemiological studies. Based on the inclusion criteria of the respective study, number of volunteers needed, potential delivery of invitation reminders, and turnaround time is set. Afterwards, matching notifications are delivered. Eventually, if volunteers autonomously decide to participate in the study, they may contact via email the promoters. Results: Between December 2020 and November 2022, the VACCELERATE Volunteer Registry has been approached to match clinical trials and epidemiological studies with volunteers 28 times. Of those, 17 (61%) studies have been focused on different aspects of SARS-CoV-2, such as finding of the best vaccination scheme, new vaccine testing, or infection point-prevalence. Additionally, new vaccinations against Streptococcus pneumoniae have been the research topic of 6 (21%) studies. Overall, 71,758 study participations have been offered to VACCELERATE volunteers of all ages. Summary/Conclusion: After 2 years since its activation, the VACCELERATE Volunteer Registry has become a reference for study sponsors and clinical trial units to speed up volunteer enrolment in trials. We expect a further increase in volunteer registration, and study requests. Keywords: BMT, Clinical trial, Vaccination, CAR-T
PB2286: EVALUATION OF THE THERAPEUTIC RESULTS OF LYMPHOBLASTIC LYMPHOMA TREATED ACCORDING TO THE EORTC 58951 PROTOCOL IN CHILDREN ADOLESCENTS AND YOUNG ADULTS
Authors: Hana Ben Hammamia, Emna Azza, Hela Sahbani, Yosr Ben Abdennabi, Zied Daoud, Raihane Ben Lakhal, Lamia Aissaoui
Journal: HemaSphere (2023)

Topic: 19. Aggressive Non-Hodgkin lymphoma - Clinical Background: Lymphoblastic lymphoma (LBL) is an aggressive neoplasm and it is one of the most common types of non-Hodgkin lymphoma (NHL) in children, adolescents and young adults (AYA). Aims: The aim of this study was to evaluate the therapeutic results in this population with LBL treated in Aziza Othmana Hospital of Tunis, Tunisia. Methods: This was a retrospective and descriptive study of 31 patients treated in Aziza Othmana hospital of Tunis from January 2009 to December 2019. The positive diagnosis was based on histological examination and immunohistochemistry. All patients were treated according to the EORTC 58951 protocol. Results: The population of our study consisted in 24 boys (77.4%) and 7 girls (22.6%) with a sex ratio of 3/1. The average age was 16 years with extremities between 5 and 25 years: 11 patients aged less than 15 years and 20 patients aged more than 15 years. Twenty-one patients (67,8%) had T-LBL and 10 patients (32,2%) had B-LBL. The tumour syndrome was the most frequent clinical presentation (61.3 %). Enlargement of the mediastinum was found in 4 patients (13%). No patient presented with testicular involvement. Four patients (12.9%) had central nervous system involvement. Among the patients studied: 9 patients (29%) were at stage II, 6 patients (19%) were at stage III and 16 patients were at stage IV (51%) according to Ann Arbor classification. Lung and pleural involvement were present in 5 patients (16.1%) each. Abdominal involvement was present in 3 patients (9.7%), kidney involvement in 1 patient (3.2%) and bone involvement in 2 patients (6.5%). Ten patients (32%) had bone marrow involvement. None of the patients presented with tumour lysis syndrome. Patients were included in a very low risk group (VLR) (1 patient), average risk 1 group (AR1) (2 patients, 6,4%), average risk 2 group (AR2) (20 patients, 64,5%) and very high-risk group (VHR) (6 patients, 19,35%). Complete remission after induction (CR) was achieved in 27patients (87%). Six patients (19%) died: 3 of them died from disease and the others died because of chemotherapy’s toxicity. We summarized the different results of five year overall survival (OS), event free survival (EFS) and free relapse survival in two groups: children’s group versus adolescents and young adults (AYA) group. - Population Number LBL-B LBL-T CR Toxic deaths OS EFS RFS <15 years 11 3 8 10(90, 9%) 1 (9%) 79,5% 58,4% 64,3% ≥15 years 20 7 13 18 (90%) 2 (10%) 79,4% 79,4% 88,2% Summary/Conclusion: The survival rates in our study were in line with literature. AYA patients had significantly better outcomes when treated with pediatric protocol. The used therapeutic protocol was associated with satisfactory results and tolerable treatment-associated side effects. Keywords: Children, Lymphoma therapy, Adolescents
PB1716: COMPARISON BETWEEN CONVENTIONAL KARYOTYPING AND INTERPHASE FISH ON AIR-DRIED SMEARS IN EVALUATING STRUCTURAL CHROMOSOMAL ABNORMALITIES AMONG PEDIATRIC ACUTE LEUKEMIA PATIENTS
Authors: Karthik Bommannan, Jhansi Rani Arumugam, Teena Koshy, Venkatraman Radhakrishnan, Shirley Sundersingh
Journal: HemaSphere (2023)

Topic: 1. Acute lymphoblastic leukemia - Biology & Translational Research Background: Leukemia-associated structural chromosomal abnormalities (SCA) can be identified either by karyotyping or interphase-fluorescence in-situ hybridization (i-FISH) assays. Both karyotyping and i-FISH on mononuclear cell suspension are time, resource and manpower consuming assays. Aims: In this study, we have compared the results of bone marrow (BM) karyotyping and i-FISH on air-dried BM/ peripheral blood (PB) smears in identifying specific leukemia-associated SCA. Methods: With Institute’s ethics committee approval, the study was conducted among pediatric patients (age <18 years) diagnosed with acute leukemias between January 2018 to December 2022. The results of i-FISH on air-dried BMA/PB smears and BM-karyotyping for our SCA of interest (BCR::ABL1, ETV6::RUNX1, TCF3::PBX1, KMT2A rearrangement, RUNX1::RUNX1T1, CBFB::MYH11, and PML::RARA) were compared. Results: Among 270 patients, SCA of interest were identified among 26% and 17% of patients by i-FISH on air-dried smears and karyotyping, respectively (Refer Table 1). Excluding 53 patients with metaphase failure, the remaining 217 patients had 92% agreement (Cramer’s V of 0.931 with p<0.000) between the results for specific SCA identified by both techniques. On excluding samples with cryptic cytogenetic aberrancies, there was 99% agreement (Cramer’s V of 0.953 with p<0.000) for gross SCA identified by both techniques. In addition, i-FISH on air-dried smears identified SCA in 30% of patients with metaphase failure. Conclusion: I-FISH on air-dried PB/BMA smears is a less-labor & resource consuming assay and can be considered as an efficient alternative for karyotyping to identify specific SCA of interest. This assay will be useful for economical evaluation of SCA by FISH in under-resourced laboratories. Table 1. Results of structural chromosomal abnormalities identified by karyotyping and interphase FISH on air-dried peripheral blood and bone marrow smears. - Karyotyping i-FISH on air-dried smears Total cases 270 270 No metaphase availability 53 Not applicable Samples reported 217 270 Positive for SCA being evaluated 36 of 217 (17%) 69 of 270 (26%) t(9;22)(q34.1;q11.2)/BCR::ABL1* 8 of 195 (4%) 13 of 246 (5%) t(;11q23.3)/KMT2A rearrangement* 3 of 192 (1.5%) 5 of 243 (2%) t(12;21)(p13.2;q22.1)/ ETV6::RUNX1* 0 of 184 (0%) 19 of 234 (8%) t(1;19)(q23;p13.3)/TCF3::PBX1* 9 of 111 (8%) 14 of 153 (9%) t(15;17)(q24;q21)/PML::RARA* 12 of 21 (57%) 13 of 23 (56.5%) inv(16)(p13.1q22) or t(16;16)(p13.1;q22)/CBFB rearrangement* 0 of 7 (0%) 0 of 8 (%) t(8;21)(q22;q22.1)/RUNX1::RUNX1T1* 4 of 7 (57%) 5 of 8 (63%) * By interphase FISH. Keywords: FISH, Acute leukemia, Cytogenetics, Pediatric